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Identification and optimization of critical process parameters for the production of NOMV vaccine against Neisseria meningitidis

机译：鉴定和优化生产针对脑膜炎奈瑟氏球菌的NOMV疫苗的关键工艺参数

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摘要

Outer membrane vesicles (OMV) are used as a vaccine against Neisseria meningitidis serogroup B and are traditionally produced with detergent-extraction to remove toxic lipopolysaccharide. Engineered strains with attenuated lipopolysaccharide allowed the use of native vesicles (NOMV) with improved stability and immunogenicity. In the NOMV production process detergents are omitted and vesicle release is stimulated with EDTA extraction (a chelating agent) to enable a higher yield. Many process parameters may change the EDTA extraction efficiency, but it is unknown what the optimal ranges for these parameters are in terms of quality. The present study systematically optimized EDTA extraction and was representative for production at large-scale. Two critical process parameters were identified, harvest point of the cultivation (harvest) and pH of the extraction buffer (pH), which significantly affected yield (7-fold) and bacterial lysis (35-fold). The other quality attributes remained unchanged. Optimization of harvest and pH revealed that the desired low bacterial lysis coincided with intermediate but sufficient yield. High functional immunogenicity and low toxicity of the optimized vaccine were also confirmed. The EDTA extraction is therefore a robust process step which produces high quality OMV if harvest and pH are controlled accurately. (C) 2012 Elsevier Ltd. All rights reserved.
机译：外膜囊泡（OMV）用作抗脑膜炎奈瑟氏菌血清群B的疫苗，传统上是用去污剂提取法生产的，以去除有毒的脂多糖。具有减脂多糖的工程菌株允许使用具有改善的稳定性和免疫原性的天然囊泡（NOMV）。在NOMV生产过程中，省去了去污剂，并通过EDTA萃取（螯合剂）刺激了囊泡的释放，从而实现了更高的收率。许多工艺参数可能会改变EDTA的萃取效率，但就质量而言，这些参数的最佳范围尚不清楚。本研究系统地优化了EDTA的提取，并具有大规模生产的代表性。确定了两个关键过程参数，即收获的收获点（收获）和提取缓冲液的pH值（pH），这显着影响产量（7倍）和细菌裂解（35倍）。其他质量属性保持不变。收获物和pH值的优化表明所需的低细菌裂解率与中等水平但足够的产量相吻合。还证实了优化疫苗的高功能免疫原性和低毒性。因此，EDTA萃取是一个可靠的工艺步骤，如果准确控制收获和pH值，则可产生高质量的OMV。 （C）2012 Elsevier Ltd.保留所有权利。

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